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Updated: Aug 8, 2026
Analysis of Physiologic E-Selectin-Mediated Leukocyte Rolling on Microvascular Endothelium
Published on: February 11, 2009
Direct real-time observation of E- and P-selectin-mediated rolling on cutaneous lymphocyte-associated antigen
Robert C Fuhlbrigge1, Sandra L King, Charles J Dimitroff
1Department of Dermatology, Brigham and Women's Hospital, Boston, MA 02115, USA.
Insights
Human memory T cells use cutaneous lymphocyte-associated antigen (CLA) on P-selectin glycoprotein ligand-1 (PSGL-1) to bind E-selectin. This study confirms CLA(+) PSGL-1 is a functional ligand for E- and P-selectin, identifying another E-selectin ligand.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Human memory T cells involved in skin inflammation express cutaneous lymphocyte-associated antigen (CLA).
- CLA is a carbohydrate determinant found on P-selectin glycoprotein ligand-1 (PSGL-1).
- CLA expression on PSGL-1 is linked to E-selectin ligand activity, but its direct role and other ligands are unclear.
Purpose of the Study:
- To determine if CLA(+) PSGL-1 is a direct ligand for E-selectin on human T cells.
- To investigate if other glycoproteins contribute to E-selectin-dependent rolling.
- To characterize the functional adhesive interactions of selectin ligands.
Main Methods:
- Developed a real-time shear flow assay to analyze cell adhesion.
- Utilized SDS-PAGE to resolve leukocyte ligands.
- Immobilized ligands on Western blots for functional analysis.
Main Results:
- Provided direct evidence that CLA(+) PSGL-1 functions as a ligand for both E-selectin and P-selectin.
- Confirmed P-selectin binding activity of PSGL-1 is not dependent on CLA modification.
- Identified a novel, non-PSGL-1 E-selectin ligand on CLA-positive human memory T cells.
Conclusions:
- CLA(+) PSGL-1 is a key functional ligand for E-selectin and P-selectin in human T cells.
- The findings elucidate the molecular mechanisms of T cell adhesion in inflammatory responses.
- A distinct E-selectin ligand, separate from PSGL-1, contributes to T cell recruitment.
Abstract:
Human memory T cells associated with cutaneous inflammatory responses are characterized by their expression of cutaneous lymphocyte-associated Ag (CLA), a carbohydrate determinant differentially expressed on P-selectin glycoprotein ligand-1 (PSGL-1). Although expression of the CLA epitope on PSGL-1 (CLA(+) PSGL-1) by memory T cells is associated with acquisition of E-selectin ligand activity, it is not known whether CLA(+) PSGL-1, itself, is a ligand for E-selectin on human T cells or whether other glycoproteins, with or without CLA modification, support E-selectin-dependent rolling in shear flow. To address this issue, we developed a method for real-time analysis of functional adhesive interactions between selectin-bearing cells in shear flow with leukocyte ligands resolved by SDS-PAGE and immobilized on standard Western blots. The results of these studies provide direct evidence that CLA(+) PSGL-1 is a functional ligand for both E- and P-selectin, confirm that the P-selectin ligand activity of PSGL-1 is independent of CLA modification, and identify a distinct, non-PSGL-1 E-selectin ligand on CLA-positive human memory T cells.
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